Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial

C. Woudstra, H. Skarin, F. Anniballi, B. Auricchio, D. De Medici, L. Bano, I. Drigo, T. Hansen, Ch. Löfström, R. Hamidjaja, B. van Rotterdam, M.G.J. Koene

Research output: Contribution to journalArticleAcademicpeer-review

5 Citations (Scopus)

Abstract

Two real-time PCR arrays based on the GeneDisc(®) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc(®) arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4%-100% for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1% to 7.1%. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.
Original languageEnglish
Pages (from-to)31-37
JournalAnaerobe
Volume22
DOIs
Publication statusPublished - 2013

Fingerprint

Clostridium botulinum type D
Clostridium botulinum type C
Dilatation and Curettage
Multicenter Studies
Real-Time Polymerase Chain Reaction
Botulism
Denmark
Sweden
Netherlands
Italy
France
Technology
Polymerase Chain Reaction
DNA

Keywords

  • quantitative detection
  • clinical-samples
  • toxin
  • gene
  • neurotoxins
  • strains
  • food

Cite this

Woudstra, C. ; Skarin, H. ; Anniballi, F. ; Auricchio, B. ; De Medici, D. ; Bano, L. ; Drigo, I. ; Hansen, T. ; Löfström, Ch. ; Hamidjaja, R. ; van Rotterdam, B. ; Koene, M.G.J. / Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial. In: Anaerobe. 2013 ; Vol. 22. pp. 31-37.
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abstract = "Two real-time PCR arrays based on the GeneDisc({\circledR}) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc({\circledR}) arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4{\%}-100{\%} for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1{\%} to 7.1{\%}. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.",
keywords = "quantitative detection, clinical-samples, toxin, gene, neurotoxins, strains, food",
author = "C. Woudstra and H. Skarin and F. Anniballi and B. Auricchio and {De Medici}, D. and L. Bano and I. Drigo and T. Hansen and Ch. L{\"o}fstr{\"o}m and R. Hamidjaja and {van Rotterdam}, B. and M.G.J. Koene",
year = "2013",
doi = "10.1016/j.anaerobe.2013.05.002",
language = "English",
volume = "22",
pages = "31--37",
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Woudstra, C, Skarin, H, Anniballi, F, Auricchio, B, De Medici, D, Bano, L, Drigo, I, Hansen, T, Löfström, C, Hamidjaja, R, van Rotterdam, B & Koene, MGJ 2013, 'Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial', Anaerobe, vol. 22, pp. 31-37. https://doi.org/10.1016/j.anaerobe.2013.05.002

Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial. / Woudstra, C.; Skarin, H.; Anniballi, F.; Auricchio, B.; De Medici, D.; Bano, L.; Drigo, I.; Hansen, T.; Löfström, Ch.; Hamidjaja, R.; van Rotterdam, B.; Koene, M.G.J.

In: Anaerobe, Vol. 22, 2013, p. 31-37.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial

AU - Woudstra, C.

AU - Skarin, H.

AU - Anniballi, F.

AU - Auricchio, B.

AU - De Medici, D.

AU - Bano, L.

AU - Drigo, I.

AU - Hansen, T.

AU - Löfström, Ch.

AU - Hamidjaja, R.

AU - van Rotterdam, B.

AU - Koene, M.G.J.

PY - 2013

Y1 - 2013

N2 - Two real-time PCR arrays based on the GeneDisc(®) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc(®) arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4%-100% for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1% to 7.1%. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.

AB - Two real-time PCR arrays based on the GeneDisc(®) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc(®) arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4%-100% for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1% to 7.1%. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.

KW - quantitative detection

KW - clinical-samples

KW - toxin

KW - gene

KW - neurotoxins

KW - strains

KW - food

U2 - 10.1016/j.anaerobe.2013.05.002

DO - 10.1016/j.anaerobe.2013.05.002

M3 - Article

VL - 22

SP - 31

EP - 37

JO - Anaerobe

JF - Anaerobe

SN - 1075-9964

ER -